1.Generation and expression of Epstein-Barr Virus Latent membrane protein 2A recombinant adenovirus
Guangyong PENG ; Kun YAO ; Lin XU
Chinese Journal of Immunology 1985;0(03):-
Objective:To generate Epstein-Barr virus(EBV) Latent Membrane Protein 2A(LMP2A) recombinant adenovirus,and provide for further investigation on the therapy vaccine against EBV associated malignancies.Methods:Full length cDNA of encoding LMP2A of EBV had been amplified by reverse transcription-PCR and cloned into pGEM-T vector.The encoding cDNA of LMP2A was inserted into E1,E3-substituted adenovirus vector pAX1CW,then the LMP2A recombinant adenovirus vector was contransfected into 293 cells togetherwith EcoT221 digested Ad5-TPC.The LMP2A recombinant adenovirus was generated by homologous recombination,and primarily identificated by ClaI enzyme digestion.The expression of LMP2A on CV1 cells infected with recombinant adenovirus analyzed by fluorescence-activated cell sorting(FACS) and confocal microscope.Results:The replication-deficient LMP2A recombinant adenovirus was generated efficiently with the titers of 2.3?10 8 pfu/ml.The LMP2A could be seen on CV1 cells membrane with confocal microscope 48 h post infected with recombinant adenovirus and the percentage of CV1 cells expressing LMP2A was 94.4% by means of FACS analysis.Conclusion:These suggested that LMP2A could be expressed efficiently by recombinant adenovirus mediated transfer,and it was the foundation of further researching in its function and developing the suitable genetic engineering vaccine against EBV associated malignancies.
2.Application of ultrasonography information system
Kun SUN ; Hong ZHANG ; Xiaojun XU ; Lin XU
Chinese Medical Equipment Journal 2004;0(07):-
This article introduces the functions and characters of a new administration system for routine pursuits in ultrasonography department. Using the framework of SAN,the system realizes equipment share and selection of storage networks according to different application condition. With powerful function and board coverage area,the administration system will positively affect the digital construction of hospitals and their departments
3.Psychometric validation of the Chinese version of Temperament Evaluation of Memphis Pisa Paris and San Di-ego-Auto questionnaire (TEMPS-A) for adolescents
Kangguang LIN ; Lingyu CHEN ; Kun CHEN ; Huiyi OUYANG ; Guiyun XU
Chinese Journal of Nervous and Mental Diseases 2016;42(6):352-356
Objective Based on the Chinese full version of the Temperament Evaluation of Memphis Pisa Paris and San Diego-Auto questionnaire ( TEMPS-A) , we aimed to validated a short version of TEMPS-A for Chinese adoles-cents.Methods Taking into account the item factor loading from our previously validated full version TEMPS-A, cultur-ally-determined items, and sexual activity related items, we derived a 60-item Chinese TEMPS-A for adolescents.The internal consistency and structural validity of the Chinese TEMPS-A for adolescents was evaluated in 822 participants aged 11~17 years old.Results The Cronbach’ s alphas coefficients for depressive, cyclothymic, hyperthymic, irritable and anxious subscales were 0.67, 0.78, 0.76, 0.77 and 0.83, respectively.The anxious, irritable, hyperthymic factors were effectively distinguished by exploratory factor analysis, while the depressive and cyclothymic factors tended to correlate.The males scored significantly higher on the hyperthymic subscale than the females [(5.407 ±2.842) vs.(4.852 ±2.963), P<0.01].The females scored significantly higher on the depressive [(3.521 ±2.221) vs.(3.144 ±2.295)], cyclothy-mic [(4.484 ±2.922) vs.(3.917 ±2.823)] and anxious [(5.236 ±3.719) vs.(4.366 ±3.658)] temperaments than the males ( P<0.05) .The scores of depressive subscale and cyclothymic temperaments subscale were significantly correla-ted (r=0.625, P<0.001), so did the scores of anxious subscale and irritable temperament subscale (r=0.628, P<0.001).Conclusions The Chinese Adolescent version of TEMPS-A is a reliable and valid instrument for investigating af-fective temperaments for adolescents.
4.Antagonistic Effect of N-acetylcysteine on Hepatic Mitochondria Damage Induced by Cadmium in Rats
Kun GUAN ; Zhao-Fa XU ; Fang-Lin ZHANG ; Al ET ;
Journal of Environment and Health 2007;0(08):-
Objective To study the antagonistic effect of N-acetylcysteine(NAC)on the damage of hepatic mitochondria of rats induced by cadmium in vitro.Methods The mitochondria were prepared from the clean Wistar rats' whole liver by using differ ential centfifugation.The mitochondria were incubated in the assay buffer containing different concentration of CdCl_2 (10,100,1 000,10 000 ?mol/L)at 37 ℃ for 1 h.The effect of NAC(500 ?mol/L)was studied at a CdCl_2 concentration of 1 000 ?mol/L.The incubation buffer was collected and the level of GSH,cytochrome C and the activity of Mn-SOD were determined. Results Compared with the control group,the level of GSH and Mn-SOD in 100,1 000,10 000 ?mol/L CdCl_2 groups were significantly decreased,the content of cytochrome C in 1 000,10 000 ?mol/L CdCl_2 groups were significantly increased(P
5.3-MA combined with TSA inhibits growth of triple-negative breast cancer cells
Kun ZHU ; Ming XU ; Mina DING ; Zhenhua LIN ; Liyan CHEN
Chinese Journal of Pathophysiology 2017;33(8):1524-1527
AIM: To investigate the effect of 3-methyladenine (3-MA) combined with trichostatin A (TSA) on triple-negative breast cancer cells and the mechanism involved.METHODS: The viability of MDA-MB-231 cells was detected by MTT assay, the migration ability was determined by scratch assay, and the expression of autophagy-related proteins was detected by Western blot.RESULTS: TSA significantly inhibited the viability of MDA-MB-231 cells in a time-and dose-dependent manner.The results of scratch assay showed that TSA inhibited cell migration ability.Western blot data indicated that TSA resulted in a moderate increase in LC3-Ⅱ expression.Moreover, 3-MA inhibited cell autophagy induced by TSA, and combination of 3-MA and TSA further inhibited the viability of MDA-MB-231 cells.CONCLUSION: Combination of 3-MA and TSA may effectively inhibit the growth of triple-negative breast cancer cells.
6.Clinical observation of 15° face-changing acetabular cup in total hip replacement for the treatment of developmental dysplasia of hip.
Wei-kun HOU ; Lin LIU ; Chao LU ; Kan PENG ; Zhi YANG ; Ke XU ; Peng XU
China Journal of Orthopaedics and Traumatology 2016;29(6):526-529
OBJECTIVETo explore early clinical effect of acetabular cup in total hip replacement for the treatment of Crowe II developmental dysplasia of hip.
METHODSEighteen patients (18 hips) with Crowe type II developmental dysplasia of hip were treated with total hip replacement from September 2001 to July 2013. Among them,including 13 males and 5 females aged from 42 to 60 years old with an average of 47.6 years old; the courses of diseases ranged from 9 to 22 years with an average of 13.5 years. All the patients had hip joint pain, limb shortening and limited hip function before operation. Harris score of hip joint were used to evaluate recovery of function at 1 day and 12 months after operation. Prosthetic coverage of acetabular cup at 1 week after operation was observed by using radiography.
RESULTSEighteen patients (18 hips) were followed up from 12 to 24 months with an average 17 months. All incisions were healed at stage I. No deep vein thrombosis, hip dislocation, periprosthetic joint infection and prosthesis loosening were occurred. No revision surgery during follow-up period. Prosthetic coverage of acetabular cup was more than 80% at 1 week after operation. Harris score were increased from 42.67 ± 5.06 before operation to 94.79 ± 3.27 at 12 months after operation (t = -45.269, P < 0.001).
CONCLUSIONFor type Crowe II developmental dysplasia of hip patients, 15° face-changing acetabular cups in THR could obtain higher actebular component coverage rate and satisfactory early clinical effects.
Acetabulum ; surgery ; Adult ; Arthroplasty, Replacement, Hip ; instrumentation ; Female ; Follow-Up Studies ; Hip Dislocation, Congenital ; surgery ; Hip Joint ; surgery ; Hip Prosthesis ; Humans ; Male ; Middle Aged
7.Preparation of polysaccharide complex and its indirect antitumor activity in vitro
Lingyan YE ; Ming REN ; Lin LYU ; Li LI ; Yanfei QI ; Juan LI ; Kun XU
Journal of Jilin University(Medicine Edition) 2014;(5):1033-1037
Objective To extract the Ginseng polysaccharide (GPS), polysaccharides of Tricholoma matsutake (PTM)and polysaccharide of Lentinus edodes (PLE)from gingeng, tricholoma matsutake and lentinus edodes respectively,and to analyze and identify their structures,and to prepare their complex,and to study the indirect antitumor activity invitro of polysaccharide complex.Methods The polysaccharides were extracted with hot water and precipitated by ethanol.The carbohydrate levels were determined by the method of phenol-sulfuric acid.The m-hydroxyphenyl method was used to determine the levels of uronic acid, and the national standard method was used to determine the levels of starch.Infrared spectroscope and chemical methods were performed to analyze their structures. Orthogonal experiment was used to study mixing methods. Cytotoxic T lymphocyte experiment and LDH release assay were performed to detect the influence of polysaccharide complex of GPS,PTM,and PLE in the CTL killing activity,and its indirect killing effect on the P815 cells.Results The extraction rates of GPS,PTM, and PLE were 8.85%,9.40%,and 10.50%;the levels of total polysaccharides were 62.96%,59.13%,and 33.86%;the levels of uronic acid were 16.44%,9.37%,and 16.44%;the starch levels were 7.26%,2.80%,and 3.77%,respectively.The identification results showed that the polysaccharides were obstrained.When the quality ratio of the three kinds of polysaccharides was 1∶1∶1 and the concentration was 600 mg·L-1 ,the CTL cytotoxicity was the highest.Conclusion The polysaccharide complex is obtained,identified and characterized. Polysaccharide complex can enhance the cytotoxicity of CTL and has the indirectly inhibitory effect on the proliferation of P815 cells.
8.Changes of 5-hydroxytryptamine and Tryptophan Hydroxylase Expressions in Tissues of Depression Model Rats Induced by Chronic Low-intensity Stress
Yi CHEN ; Haixia XU ; Mingyue ZHU ; Kun LIU ; Bo LIN ; Ruxian LUO ; Chuanbai CHEN ; Mengsen LI
China Pharmacy 2017;28(22):3061-3064
OBJECTIVE:To provide basis and thinking for the development of medicines for treating depression. METHODS:Rats with adaptive feeding for 7 d were randomly divided into normal group,depression group and treatment group(citalopram hy-drobromide 1 mg/kg),8 in each group. Rats in depression group and treatment groups randomly received once low-density stimula-tion in a day [catching tail stimulation(1 min),frequent flashing stimulation(120 times/min,12 h),noise stimulation(4 h),ice water swimming stimulation(1 h),hot water stimulation(45 ℃,1 h),electric shocking foot(10 Hz,1 mA,each time interval of 100 ms,100 times),no food and no water(24 h)] for 21 d to reduce stress depression model. Body mass of rats in 0,7 d of adaptive feeding and 7,14,21 d of stress were respectively weighed,and horizontal activity and vertical activity frequencies in open field test were detected. After stress,rats were intragastrically administrated related medicines for 30 d,then 5-hydroxytrypta-mine (5-HT) and tryptophan hydroxylase (TPH) expressions in brain,liver,kidney tissues were determined. RESULTS:Com-pared with normal group,the increasing trend of body mass in depression group and treatment group slowed down from the 14 d of stress(P<0.05). The horizontal activity frequencies in depression group and treatment group in 14,21 d of stress slowed more ob-viously than the previous time point (P<0.05 or P<0.01);vertical activity frequencies in 21 d of stress slowed more obviously than the previous time point (P<0.05 or P<0.01). After 30 d of administration,horizontal activity and vertical activity frequen-cies in treatment group were increased more obviously than the previous time point (P<0.01). Compared with depression group,brain tissue of rats,indicating that TPH can be used as a target for depression treatment.
9.Roles of CD4 + CD25 + regulatory T cells and T helper 17 cells in the pathogenesis of Kawasaki disease
Zhao DUAN ; Zhiping LIN ; Qing LIU ; Kun XU ; Zhiqiang LIU ; Zheng ZOU
Chinese Journal of Applied Clinical Pediatrics 2017;32(9):652-655
Objective To observe the changes in CD4 + CD25 + regulatory T cells(Treg) and T helper 17 cells (Th17) cells' proportions in the peripheral blood in children with Kawasaki disease(KD) before and after the treatment,and to analyze the role of Treg/Th17 cell imbalance in the pathogenesis of KD.Methods Fifty-two children with acute KD(KD group) and 34 age-matched healthy children(healthy control group) were selected at Jiangxi Provincial Children's Hospital from April to December of 2014.Morning peripheral vein blood was collected from 2 groups:one before the treatment by Immunoglobulin and Aspirin,and the other 3 days after defervescence treatment.Flow cytometry was used to detect proportions of Treg cells and Th17 cells in the peripheral blood.The enzyme linked immunosorbent assay was used to detect the levels of interleukin(IL)-6,IL-10,IL-17,IL-23 and transforming growth factor(TGF)-β.Results Proportion of Treg cells in the acute KD group was remarkably lower than that in the healthy control group [(1.48 ± 0.21) % vs.(5.13-± 0.32) %,t =28.41,P < 0.05],but it was significantly increased after treatment,and there was a significant difference [(4.71 ± 0.36) % vs.(1.48 ± 0.21) %,t =-23.32,P < 0.05].Proportion of Th17 cells in the acute KD group was markedly higher than that in the healthy control group [(8.06 ± 0.48) % vs.(2.65 ± 0.50) %,t =-23.47,P < 0.05],which was significantly decreased after treatment [(3.04 ±0.35) % vs.(8.06 ± 0.48) %,t =25.55,P < 0.05].Compared with the healthy control group,the levels of serum IL-6,IL-17,IL-23 in the acute KD group were significantly increased before treatment,and there were significant differences [(34.53 ± 0.53) ng/L vs.(10.88 ± 0.83) ng/L,t =-72.36;(57.05 ± 0.78) ng/L vs.(14.29 ± 0.98)ng/L,t =-55.29;(45.18 ± 1.52) ng/L vs.(18.25 ± 1.08) ng/L,t =-43.27;all P < 0.05],but after treatment the levels were significantly decreased [(14.94 ± 1.06) ng/L vs.(34.53 ± 0.53) ng/L,t =49.63;(27.64 ± 0.91)ng/Lvs.(57.05±0.78) ng/L,t =26.49;(24.50-±1.13) ng/L vs.(45.18-±1.52) ng/L,t =32.17;allP<0.05].The levels of serum IL-10,TGF-β in the acute KD group significantly decreased than those of the healthy control group,and there were significant differences [(14.29-± 0.64) ng/L vs.(29.57 ± 0.87) ng/L,t =42.24;(16.88 ±-0.90) ng/L vs.(38.83 ±0.84) ng/L,t =53.51;all P <0.05],but after treatment the levels were significantly increased,and there were significant differences [(23.01-± 0.61) ng/L vs.(14.29-± 0.64) ng/L,t =-29.54;(33.47±-0.82) ng/Lvs.(16.88±-0.90) ng/L,t=-40.68;allP<0.05].Conclusion Imbalance betweenTreg cells and Th17 cells may be an important cause for the immune disorder of KD,the changes in related cytokines are involved in the pathogenesis of KD.
10.Clinical analysis of laparoscopic cholecystectomy and its complications
Kun OU ; Qiang-Pu CHEN ; Qing-Hai GUAN ; Fan ZHANG ; Xu-Tao LIN ;
Chinese Journal of Primary Medicine and Pharmacy 2006;0(08):-
Objective To explore the prevention and treament of the complications for laparoscopic cholecys- teetomy(LC).Methods Clinical datas of 600 eases undergoing LC were collected and analyzed retrospectively.Re- suits Among all the cases,249 cases were diagnosed as cholecystolithiasis,284 cases as gallbladder and 67 cases as chronic cholecystitis.The mean duration of operation was 42min.Conversions to open cholecystectomy were needed in 18 cases(3%).Complications took place in 6 cases(0.5%),involving 2 cases of abdominal bleeding and 1 case of bile leakage.There was no bile duct injury and death.Conclusion Strictly-controlled indications,skilled performance and timely open exploration are very important to prevent and reduce the complications.